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Photo-controllable ATP (Adenosine Triphosphate) biosynthesis system and preparation method thereof
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A biosynthesis, system technology, applied in the direction of fermentation, etc.
Active Publication Date: 2014-09-24
INST OF CHEM CHINESE ACAD OF SCI
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So far, there is no report of using photosystem II protein as proton source to promote ATP synthesis
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[0045] (b) Disperse calciumcarbonate microspheres in MES (2-(N-morpholine)ethanesulfonic acid) solution containing 0.025wt% GA (glutaraldehyde), cross-link for 3 hours, wash by centrifugation, and remove with 0.1M EDTA Template calcium carbonate particles; continue to disperse in MES solution containing 0.3mg chl / mL photosystem II, cros...
Embodiment 2
[0055] Embodiment 2, preparation photosystem II / ATP synthase assembly system
[0056] (1) Preparation of photosystem II microcapsules
[0057] Manganese carbonate particles with a particle size of 5 microns were dispersed in a solution containing 0.2M sodiumchloride and 2 mg / mL polyacrylamide for 30 minutes, then centrifuged, washed thoroughly, and then dispersed in a solution containing 0.025 wt% GA. React for 3 hours, wash thoroughly, and then disperse into 40mmol MES buffer solution (pH=6) containing 0.5mg / mL photosystem II and 2mg / mL BSA, shake and adsorb for 4h, the aldehyde group on GA reacts with the amino group of the protein, thereby The protein is immobilized on the surface of the particle, and an assembly cycle is completed, and the operation of adsorbing GA and photosystem II is repeated in sequence until the required number of layers. The template manganese carbonate particles were removed with 0.1M EDTA to obtain BSA / PSII / GA covalently cross-linked microcapsule...
Embodiment 3
[0062] Embodiment 3, preparation photosystem II / ATP synthase assembly system
[0063] (1) Preparation of photosystem II microspheres
[0064] Disperse silica particles with a particle size of 500nm in a solution of 1mg / mL polyacrylamide containing 0.2M sodiumchloride and absorb for 30min, then centrifuge, wash thoroughly, and then disperse in a solution containing 0.025wt% GA, React for 3 hours, wash thoroughly, and then disperse into 40mmol MES buffer solution (pH=6) containing 0.5mg / mL Photosystem II and 2mg / mL BSA, shake and adsorb for 4h, completing one assembly cycle, and repeat the adsorption of GA and photosystem II in sequence. System II operates until the desired number of layers. Nano-silica particles are biocompatible, have no effect on protons, and do not need to remove nuclei. Microspheres with a diameter of about 500nm were obtained. The beads were dispersed in buffer and stored at 4°C.
[0065] (2) The preparation of proteoliposomes containing ATP synthase ...
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Abstract
The invention discloses a photo-controllable ATP (Adenosine Triphosphate) biosynthesissystem and a preparation method thereof. The method comprises the following steps: (1) preparing photosystem II microspheres or photosystem II capsules; (2) dispersing the photosystem II microspheres or the photosystem II capsules in a 2-(N-morpholine) ethanesulfonic acidbuffer solution containing a dialdehyde cross-linking agent for adsorption; next, continuing dispersing the photosystem II microspheres or the photosystem II capsules in the 2-(N-morpholine) ethanesulfonic acidbuffer solution containing the photosystem II for adsorption; (3) dispersing the photosystem II microspheres or the photosystem II capsules treated previously in the step (2) in a proteoliposome solution containing ATP synthetase for adsorption, thereby obtaining the photo-controllable ATP biosynthesissystem. In the photo-controllable ATP biosynthesissystem, the photosystem II is immobilized on three-dimensional (micro / nano-spheres and micro / nano-capsules) surfaces so that the action area is increased and the applicability is improved; the dimension, the structure and the functions can be regulated and controlled; according to the actual requirement, the dimension and the structure of the spheres or the capsules can be changed optionally by selecting different synthesis methods, and the spheres or the capsules can be coated with extra functional capsule wall components.
Description
technical field [0001] The invention relates to an ATP synthesis system and a preparation method thereof, in particular to a light-controllable ATP biosynthesis system and a preparation method thereof. Background technique [0002] ATP is the "energy currency" in organisms and plays an important role in energy metabolism. It provides energy for a variety of energy-consuming processes in cells and participates in the regulation of various biochemical processes, such as nerve conduction, metabolism, muscle contraction, bioluminescence, and material transport. Due to its diverse functions, if the controllable synthesis of ATP in cells can be simulated at the subcellular level, it will be able to achieve fine control and research on various biological processes. [0003] ATP is synthesized by ATP synthase in living organisms. Using ATP synthase to construct the assembly structure of the biomimetic system can realize the fine control of ATP synthesis on the artificial carrier, ...
Claims
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